Objective: We performed single-cell RNA sequencing (scRNA-seq) of enriched glomerular and nonglomerular cells from 20-week-old db/m and db/db mice to generate transcriptome of 59,382 cells. To enrich glomerular cells, the kidneys were perfused with magnetic beads, and magnetic separator collected glomerular cells from dissociated kidney samples. Methods: All major cell types of the kidney were identified in the final dataset. On the basis of gene expression patterns, we subclustered podocytes and parietal epithelial cells (PECs) into 5 states consisted of healthy podocytes, stressed podocytes, dedifferentiated podocytes, transitional cells, and PECs. Cell trajectory analysis showed that a subset of PECs undergo transition into podocytes putatively regulated by combinatorial reshaping of basic leucine zipper transcription factor from Atf3, Jun, and Fos to Mafb and Xbp1. In db/db mice, podocyte regeneration was not apparent, and the majority of podocytes undergo dedifferentiation driven by upregulation of Hnf4a and Srebf2. Gene set enrichment analysis revealed that this transcriptional reprogramming is responsible for enhanced fatty acid oxdiation, gluconeogenesis, and increased oxidative phosphorylation (OXPHOS).   Proximal tubular cells showed more heterogeneous cell populations in db/db mice including immune marker-expressing cells and angiotensinogen-expressing cells. In addition, proximal tubular cells of db/db mice show distinct metabolic alteration characterized by impaired fatty acid oxidation, concomitantly increased gluconeogensis and glycolysis, and decreased OXPHOS, which are considerably different features compared to podocytes. Results: scRNA-seq of mouse kidneys revealed that transcriptional remodeling occurs in podocyte regeneration and dedifferentiation. In db/db mice, podocytes undergo dedifferentiation associated with increased fatty acid oxidation and OXPHOS, while proximal tubular cells have defective fatty acid oxidation and impaired OXPHOS. Conclusions: Objective: Diabetic kidney disease is manifested as albuminuria and progressive decline of glomerular filtration rate, which represent constellation of cellular damage in glomerular and tubular cells. The complex nature of diabetic kidney disease warrants cell type-specific molecular characterization to better understand its pathophysiology. Methods: We performed single-cell RNA sequencing (scRNA-seq) of enriched glomerular and nonglomerular cells from 20-week-old db/m and db/db mice to generate transcriptome of 59,382 cells. To enrich glomerular cells, the kidneys were perfused with magnetic beads, and magnetic separator collected glomerular cells from dissociated kidney samples. Results: All major cell types of the kidney were identified in the final dataset. On the basis of gene expression patterns, we subclustered podocytes and parietal epithelial cells (PECs) into 5 states consisted of healthy podocytes, stressed podocytes, dedifferentiated podocytes, transitional cells, and PECs. Cell trajectory analysis showed that a subset of PECs undergo transition into podocytes putatively regulated by combinatorial reshaping of basic leucine zipper transcription factor from Atf3, Jun, and Fos to Mafb and Xbp1. In db/db mice, podocyte regeneration was not apparent, and the majority of podocytes undergo dedifferentiation driven by upregulation of Hnf4a and Srebf2. Gene set enrichment analysis revealed that this transcriptional reprogramming is responsible for enhanced fatty acid oxdiation, gluconeogenesis, and increased oxidative phosphorylation (OXPHOS).   Proximal tubular cells showed more heterogeneous cell populations in db/db mice including immune marker-expressing cells and angiotensinogen-expressing cells. In addition, proximal tubular cells of db/db mice show distinct metabolic alteration characterized by impaired fatty acid oxidation, concomitantly increased gluconeogensis and glycolysis, and decreased OXPHOS, which are considerably different features compared to podocytes. Conclusions: scRNA-seq of mouse kidneys revealed that transcriptional remodeling occurs in podocyte regeneration and dedifferentiation. In db/db mice, podocytes undergo dedifferentiation associated with increased fatty acid oxidation and OXPHOS, while proximal tubular cells have defective fatty acid oxidation and impaired OXPHOS.